Demonstration by mass spectrometry that pseudo-hevein and hevein have ragged C-terminal sequences

Um, Soedjanaatmadja; Hofsteenge, Jan; Cm, Jeronimus-Stratingh; Ap, Bruins; Beintema, Jj

doi:10.1016/0167-4838(94)90150-3

Cited by 38 publications

(29 citation statements)

References 12 publications

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“…The stoichiometry (g) was also nearly one in both cases. Taken together, these results indicate that the residues in the sugar binding site, essentially Trp 21 , Trp 23 , Tyr 30 and Ser 19 , are spatially arranged in a similar way in both isoallergens [10], giving approximately the same affinity for the chitotriose moiety. Interestingly, Asensio et al [31] have also demonstrated, using nuclear magnetic resonance (NMR) techniques that the interactions of hevein with (GlcNAc) 5 oligomers involve, not only the residues indicated above, but also the loop region (Pro 13 , Lue 16 ) and Cys 24 .…”

Section: Purification and Characterization Of Hev B 60202mentioning

confidence: 63%

“…1B). In contrast, using X-ray crystallography and chemical modification of Trp 21 and Trp 23 , we determined that these aromatic residues were relevant for the interaction with model antibodies and IgE [18]. Besides, the relevance that aromatic and polar side chains might have in IgE recognition by hevein-like domains was made apparent from a primary-structure comparison including 60 domains found in proteins that are recognized as allergens (Hev b 6.02, Hev b 11, wheat germ agglutinin, WGA) or potential plant allergens belonging to the pathogenesis-related family.…”

Section: Purification and Characterization Of Hev B 60202mentioning

confidence: 99%

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A single amino acid substitution on the surface of a natural hevein isoform (Hev b 6.0202), confers different IgE recognition

et al. 2006

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Decreased immune reactivity of isoforms of major allergens has been reported. However, such claims have always been based on experiments with recombinant proteins. This work describes the molecular and physicochemical characterization of a hevein (Hev b 6.0201) natural isoform (Hev b 6.0202), which is present in rubber latex from Hevea brasiliensis. The isoallergen has a single substitution Asn14Asp, which gives rise to local differences in the surface potential, as observed from the crystal structure presented here. Besides, ELISA inhibition using serum pools of adult and pediatric patients showed reduced IgE-binding capacity ($27%) with the isoallergen. Overall, these results are relevant to delineate crucial residues involved in this dominant discontinuous epitope.

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Section: Purification and Characterization Of Hev B 60202mentioning

confidence: 63%

Section: Purification and Characterization Of Hev B 60202mentioning

confidence: 99%

A single amino acid substitution on the surface of a natural hevein isoform (Hev b 6.0202), confers different IgE recognition

et al. 2006

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“…Peptides were isolated by reverse-phase HPLC and N-terminal amino acid sequences were determined by manual Edman degradation [6]. The C-terminal peptide of the 14 kDa protein was identified by comparing its N-terminal sequence with the translated cDNA sequence of the 14 kDa protein.…”

Section: Resultsmentioning

confidence: 99%

“…N-Terminal sequence analysis of the C-terminal domain indicated that 6 amino acids between the hevein and the C-terminal domain are lost during processing of the precursor [5]. Analysis by ionspray mass spectrometry of hevein showed that minor components with one and two additional C-terminal amino acids occur [6], indicating that processing probably occurs by cleavage at the N-terminus of the C-terminal domain, followed by removal of 4-6 residues from the C-terminus of hevein [6].…”

Section: Introductionmentioning

confidence: 99%

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Processed products of the hevein precursor in the latex of the rubber tree (Hevea brasiliensis)

1995

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The 20 kDa precursor of hevein and its C-terminal 14 kDa domain have been isolated. Sequence analysis of the Cterminal tryptic peptides of these proteins and comparison with the cDNA sequence indicate that they represent mature forms from which a C-terminal propeptide, possibly involved in vacuolar targeting, has been removed. The molar ratio of hevein to the C-terminal domain in the lutoid-body fraction of rubber latex is about 30:1. This indicates that not only the pre-and propeptides but also the 14 kDa domain are removed by proteolysis or other processes in the latex vessel after the processing of hevein has taken place.the hevein domain in this protein is not cleaved off in the mature form.Here we report the isolation of the hevein precursor and its C-terminal domain and present evidence that they are the mature proteins from which a C-terminal propeptide has already been removed. The large molar excess of hevein in the lutoidbody fraction of rubber latex indicates that much proitein turnover occurs in the latex vessels of the rubber tree. Materials and methods

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Role of aromatic amino acids in carbohydrate binding of plant lectins: Laser photo chemically induced dynamic nuclear polarization study of hevein domain-containing lectins

et al. 1997

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Carbohydrate recognition by lectins often involves the side chains of tyrosine, tryptophan, and histidine residues. These moieties are able to produce chemically induced dynamic nuclear polarization (CIDNP) signals after laser irradiation in the presence of a suitable radical pair-generating dye. Elicitation of such a response in proteins implies accessibility of the respective groups to the light-absorbing dye. In principle, this technique is suitable to monitor surface properties of a receptor and the effect of ligand binding if CIDNP-reactive amino acids are affected. The application of this method in glycosciences can provide insights into the protein-carbohydrate interaction process, as illustrated in this initial study. It focuses on a series of N-acetylglucosamine-binding plant lectins of increasing structural complexity (hevein, pseudohevein, Urtica dioica agglutinin and wheat germ agglutinin and its domain B), for which structural NMR- or X-ray crystallographic data permit a decision of the validity of the CIDNP method-derived conclusions. On the other hand, the CIDNP data presented in this study can be used for a rating of our molecular models of hevein, pseudohevein, and domain B obtained by various modeling techniques. Experimentally, the shape and intensity of CIDNP signals are determined in the absence and in the presence of specific glycoligands. When the carbohydrate ligand is bound, CIDNP signals of side chain protons of tyrosine, tryptophan, or histidine residues are altered, for example, they are broadened and of reduced intensity or disappear completely. In the case of UDA, the appearance of a new tryptophan signal upon ligand binding was interpreted as an indication for a conformational change of the corresponding indole ring. Therefore, CIDNP represents a suitable tool to study protein-carbohydrate interactions in solution, complementing methods such as X-ray crystallography, high-resolution multidimensional nuclear magnetic resonance, transferred nuclear Overhauser effect experiments, and molecular modeling.

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Demonstration by mass spectrometry that pseudo-hevein and hevein have ragged C-terminal sequences

Cited by 38 publications

References 12 publications

A single amino acid substitution on the surface of a natural hevein isoform (Hev b 6.0202), confers different IgE recognition

A single amino acid substitution on the surface of a natural hevein isoform (Hev b 6.0202), confers different IgE recognition

Processed products of the hevein precursor in the latex of the rubber tree (Hevea brasiliensis)

Role of aromatic amino acids in carbohydrate binding of plant lectins: Laser photo chemically induced dynamic nuclear polarization study of hevein domain-containing lectins

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